POC

Function

To design a input (ligand) with 2 fluorescein molecules attached by a piece of DNA, which will be used in the intermediate step of testing if binding of an antigen to our system can cause dimerization and subsequent transcription of the desired output gene.

Receiver 2:

Input here is simply just a fluorecein molecule to test if binding of an antigen to our system can cause a conformation change and subsequent transcription of the desired output gene.

Device Depiction

Fluorescein Structure

Device Parts

Lengths:

10 bp
12 bp
15 bp
20 bp
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A pair of ssDNA strands complementary to each other, each bound to a fluorescein molecule at the 5' end, of the following lengths

Current Status

Completed Work

Determined the length and content of DNA between fluorescein molecules

10 bp ~ 33.2 A: because DNA twists every 33.2 A, at this point, the fluorescein molecules, on different strands of DNA, will be pointing directly away from each other, 180 degrees apart

Outer membrane: This lipid bilayer is found in Gram negative bacteria and is the source of lipopolysaccharide (LPS). LPS is toxic and turns on the immune system; LPS is found in Gram negative, but not in Gram positive, bacteria.

Cell wall is composed of peptidoglycan (polysaccharides + protein), the cell wall maintains the overall shape of a bacterial cell. The three primary shapes in bacteria are coccus (spherical), bacillus (rod-shaped) and spirillum (spiral). Mycoplasma are bacteria that have no cell wall and therefore have no definite shape.

Periplasm: This cellular compartment is found only in those bacteria that have both an outer membrane and plasma membrane (e.g. Gram negative bacteria). In the space are enzymes and other proteins that help digest and move nutrients into the cell.

Inner membrane: also known as plasma membrane. This is a lipid bilayer much like the cytoplasmic (plasma) membrane of other cells. There are numerous proteins moving within or upon this layer that are primarily responsible for transport of ions, nutrients and waste across the membrane.

Problems

The outer membrane is the major permeability barrier in Gram negative bacteria. Gram negative bacteria store degradative enzymes in the periplasmic space.

Solutions

Work with E.Coli strain that is cell-wall deficient

Or work with E.Coli strain that has the most permeable cell wall <-- LIMITATION of antigen size. Ways to make cell wall permeable are: treatment with chemicals (example: EDTA)